All-Optical Wide-Field Selective Imaging of Fluorescent Nanodiamonds in Cells, In Vivo and Ex Vivo
| Autor: | Takeshi Ohshima, Tatsuhiko Imaoka, Takuma Sugi, Hiroshi Abe, Wataru Kada, Osamu Hanaizumi, Hiroki Yamamoto, Shizuko Kakinuma, Ryuji Igarashi, Akihiro Kuwahata, Kiichi Kaminaga, Masaki Sekino, Tamami Yanagi, Michiyo Suzuki |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
Fluorescence-lifetime imaging microscopy Materials science Nitrogen Biomolecule Optical Imaging General Engineering General Physics and Astronomy Laser Fluorescence Rats Nanodiamonds law.invention Microsecond Autofluorescence chemistry law In vivo Biophysics Animals General Materials Science Coloring Agents Nanodiamond Fluorescent Dyes |
| Zdroj: | ACS Nano. 15:12869-12879 |
| ISSN: | 1936-086X 1936-0851 |
| DOI: | 10.1021/acsnano.0c07740 |
| Popis: | Fluorescence imaging is a critical tool to understand the spatial distribution of biomacromolecules in cells andiin vivo/i, providing information on molecular dynamics and interactions. Numerous valuable insights into biological systems have been provided by the specific detection of various molecular species. However, molecule-selective detection is often hampered by background fluorescence, such as cell autofluorescence and fluorescence leakage from molecules stained by other dyes. Here we describe a method for all-optical selective imaging of fluorescent nanodiamonds containing nitrogen-vacancy centers (NVCs) for wide-field fluorescence bioimaging. The method is based on the fact that the fluorescence intensity of NVCs strictly depends on the configuration of ground-state electron spins, which can be controlled by changing the pulse recurrence intervals of microsecond excitation laser pulses. Therefore, by using regulated laser pulses, we can oscillate the fluorescence from NVCs in a nanodiamond, while oscillating other optical signals in the opposite phase to NVCs. As a result, we can reconstruct a selective image of a nanodiamond by using a series of oscillated fluorescence images. We demonstrate application of the method to the selective imaging of nanodiamonds in live cells, in microanimals, and on a hippocampal slice culture obtained from a rat. Our approach potentially enables us to achieve high-contrast images of nanodiamond-labeled biomolecules with a signal-to-background ratio improved by up to 100-fold over the standard fluorescence image, thereby providing a more powerful tool for the investigation of molecular dynamics in cells andiin vivo/i. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|